The botulinum neurotoxins, BoNTs, produced by Clostridium botulinum are among the most potent toxins known to man. The Centers for Disease Control and Prevention (CDC) has classified it as a potential bio-weapon, Category A, because of its extreme potency and lethality, its ease of production and transport, and the need for prolonged intensive care among affected persons. This project focuses on generating monoclonal antibodies to be used in the development of diagnostic assays and devices and to be used as a general tool for understanding toxin structure-function relationships. The goal is to generate mAbs that recognize all neurotoxin subtypes multiple serotypes. Studies will be conducted to understand the role of antibody epitopes on toxin recognition and to use this knowledge to engineer mAbs allowing more sensitive toxin detection or greater therapeutic efficacy. Antibodies will be generated against the heavy chain domains as part of a collaboration with Project 3.1 with the aim of understanding the receptor recognition and binding process and in collaboration with Project 3.3 with the aim of understanding the translocation process. mAbs against the light chain domain will be generated for use in assays and diagnostic devices. Samples generated will be made available to PSWRCE collaborators as well as to others in the biodefense consortium.